Publications by authors named "Yongmei Xi"

30 Publications

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The S100A10-AnxA2 complex is associated with the exocytosis of hepatitis B virus in intrauterine infection.

Lab Invest 2021 Oct 13. Epub 2021 Oct 13.

The Women's Hospital, Zhejiang University School of Medicine, No. 1 Xueshi Road, Shangcheng District, Hangzhou, Zhejiang, 310001, China.

Mother-to-child transmission (MTCT) is the major cause of chronic infection of hepatitis B virus (HBV) in patients. However, whether and how HBV crosses the placenta to cause infection in utero remains unclear. In this study, we investigate the mechanism as to how the HBV virions pass through layers of the trophoblast. Our data demonstrate the exocytosis of virions from the trophoblast after exposure to HBV where the endocytosed HBV virions co-localized with an S100A10/AnxA2 complex and LC3, an autophagosome membrane marker. Knockdown of either AnxA2 or S100A10 in trophoblast cells led to a reduction of the amount of exo-virus in Transwell assay. Immunohistochemistry also showed a high expression of AnxA2 and S100A10 in the placental tissue samples of HBV-infected mothers with congenital HBV-positive infants (HBV). We conclude that in HBV intrauterine infection and mother-to-child transmission, a proportion of HBV hijacks autophagic protein secretion pathway and translocate across the trophoblast via S100A10/AnxA2 complex and multivesicular body (MVB)-mediated exocytosis. Our study provides a potential target for the interference of the mechanisms of HBV intrauterine infection and mother-to-child transmission.
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http://dx.doi.org/10.1038/s41374-021-00681-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8512653PMC
October 2021

Transcriptome-Based Analysis Reveals Therapeutic Effects of Resveratrol on Endometriosis in aRat Model.

Drug Des Devel Ther 2021 29;15:4141-4155. Epub 2021 Sep 29.

The Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310006, People's Republic of China.

Introduction: Endometriosis (EMs) is associated with severe chronic pelvic pain and infertility and the development of improved EMs treatment options is an ongoing focus. In this study, we investigated the effects of resveratrol on EMs and analyzed transcriptional changes in the lesions of model rats before and after resveratrol treatment.

Methods: We established arat model of endometriosis through the trans-implantation of endometrial fragments to the peritoneal wall and then used resveratrol as treatment. We then analyzed the results using RNA sequencing of the lesion tissues of each of the model rats before resveratrol treatment and the reduced lesion tissues after the treatment. Examinations of anatomy, biochemistry, immunohistochemical staining and flow cytometry examinations were also conducted. Other trans-implanted rats were also given sham treatments as sham-treatment control and other untrans-implanted rats served as sham-operation controls.

Results: In addition to the obvious lesions observed in the model rats, there were significant differences in the glucose tolerance, macrophage M1/M2 polarization, and adipocyte sizes between the treated model rats and sham (control) rats. Resveratrol treatment in the model rats showed significant efficacy and positive therapeutic effect. Transcriptional analysis showed that the effects of resveratrol on the endometriosis model rats were manifested by alterations in the PPAR, insulin resistance, MAPK and PI3K/Akt signaling pathways. Correspondingly, changes in PPARγ activation, M1/M2 polarization and lipid metabolism were also detected after resveratrol treatment.

Discussion: Our study revealed that resveratrol treatment displayed efficient therapeutic effects for EMs model rats, probably through its important roles in anti-inflammation, immunoregulation and lipid-related metabolism regulation.
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http://dx.doi.org/10.2147/DDDT.S323790DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8487867PMC
September 2021

Lipidomic Alterations and PPAR Activation Induced by Resveratrol Lead to Reduction in Lesion Size in Endometriosis Models.

Oxid Med Cell Longev 2021 11;2021:9979953. Epub 2021 Sep 11.

The Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, China.

Endometriosis is an estrogen-dependent chronic inflammatory disease that affects approximately 10% of women of reproductive age and up to 50% of women with infertility. The heterogeneity of the disease makes accurate diagnosis and treatment a clinical challenge. In this study, we generated two models of endometriosis: the first in rats and the second using human ectopic endometrial stromal cells (HEcESCs) derived from the lesion tissues of endometriosis patients. We then applied resveratrol to assess its therapeutic potential. Resveratrol intervention had significant efficacy to attenuate lesion size and to rectify aberrant lipid profiles of model rats. Lipidomic analysis revealed significant lipidomic alterations, including notable increases of sphingolipids and decreases of both glycerolipids and most phospholipids. Upon resveratrol application, both proliferation capacity and invasiveness parameters decreased, and the early apoptosis proportion increased for HEcESCs. The activation of PPAR was also noted as a factor potentially contributing to recovery from endometriosis in both models. Our study provides valuable insight into the mechanisms of resveratrol in endometriosis and therefore strengthens the potential for optimizing resveratrol treatment for this disease.
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http://dx.doi.org/10.1155/2021/9979953DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8452402PMC
September 2021

Premigratory neural crest stem cells generate enteric neurons populating the mouse colon and regulating peristalsis in tissue-engineered intestine.

Stem Cells Transl Med 2021 Jun 22;10(6):922-938. Epub 2021 Jan 22.

Institute of Translational Medicine, and Children's Hospital Affiliated and Key Laboratory of Diagnosis and Treatment of Neonatal Diseases of Zhejiang Province, the Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, People's Republic of China.

Hirschsprung's disease (HSCR) is a common congenital defect. It occurs when bowel colonization by neural crest-derived enteric nervous system (ENS) precursors is incomplete during the first trimester of pregnancy. Several sources of candidate cells have been previously studied for their capacity to regenerate the ENS, including enteric neural crest stem cells (En-NCSCs) derived from native intestine or those simulated from human pluripotent stem cells (hPSCs). However, it is not yet known whether the native NCSCs other than En-NCSCs would have the potential of regenerating functional enteric neurons and producing neuron dependent motility under the intestinal environment. The present study was designed to determine whether premigratory NCSCs (pNCSCs), as a type of the nonenteric NCSCs, could form enteric neurons and mediate the motility. pNCSCs were firstly transplanted into the colon of adult mice, and were found to survive, migrate, differentiate into enteric neurons, and successfully integrate into the adult mouse colon. When the mixture of pNCSCs and human intestinal organoids was implanted into the subrenal capsule of nude mice and grown into the mature tissue-engineered intestine (TEI), the pNCSCs-derived neurons mediated neuron-dependent peristalsis of TEI. These results show that the pNCSCs that were previously assumed to not be induced by intestinal environment or cues can innervate the intestine and establish neuron-dependent motility. Future cell candidates for ENS regeneration may include nonenteric NCSCs.
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http://dx.doi.org/10.1002/sctm.20-0469DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8133337PMC
June 2021

[Research progress on biomarkers for endometriosis based on lipidomics].

Zhejiang Da Xue Xue Bao Yi Xue Ban 2020 Dec;49(6):779-784

Institute of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.

The pathogenesis of endometriosis is not well understood at the moment, and the lack of effective biomarkers often leads to delayed diagnosis of the disease. Lipidomics provides a new approach for the diagnosis and prediction of endometriosis. Sphingomyelin, phosphatidylcholine and phosphatidylserine in peripheral blood, endometrial fluid, peritoneal fluid and follicular fluid have good diagnostic value for endometriosis and disease classification; the lipid metabolites in the eutopic endometrium tissue are expected to be biomarkers of early endometriosis; and the lipid metabolites in peripheral blood are also of great value for predicting endometriosis-related infertility. The development of lipidomics technique will further advance the progress on the pathogenesis, prediction, diagnosis and treatment of endometriosis.
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http://dx.doi.org/10.3785/j.issn.1008-9292.2020.12.14DOI Listing
December 2020

KVarPredDB: a database for predicting pathogenicity of missense sequence variants of keratin genes associated with genodermatoses.

Hum Genomics 2020 12 7;14(1):45. Epub 2020 Dec 7.

Department of Human Genetics, and Women's Hospital, Zhejiang University School of Medicine, Hangzhou, China.

Background: Germline variants of ten keratin genes (K1, K2, K5, K6A, K6B, K9, K10, K14, K16, and K17) have been reported for causing different types of genodermatoses with an autosomal dominant mode of inheritance. Among all the variants of these ten keratin genes, most of them are missense variants. Unlike pathogenic and likely pathogenic variants, understanding the clinical importance of novel missense variants or variants of uncertain significance (VUS) is the biggest challenge for clinicians or medical geneticists. Functional characterization is the only way to understand the clinical association of novel missense variants or VUS but it is time consuming, costly, and depends on the availability of patient's samples. Existing databases report the pathogenic variants of the keratin genes, but never emphasize the systematic effects of these variants on keratin protein structure and genotype-phenotype correlation.

Results: To address this need, we developed a comprehensive database KVarPredDB, which contains information of all ten keratin genes associated with genodermatoses. We integrated and curated 400 reported pathogenic missense variants as well as 4629 missense VUS. KVarPredDB predicts the pathogenicity of novel missense variants as well as to understand the severity of disease phenotype, based on four criteria; firstly, the difference in physico-chemical properties between the wild type and substituted amino acids; secondly, the loss of inter/intra-chain interactions; thirdly, evolutionary conservation of the wild type amino acids and lastly, the effect of the substituted amino acids in the heptad repeat. Molecular docking simulations based on resolved crystal structures were adopted to predict stability changes and get the binding energy to compare the wild type protein with the mutated one. We use this basic information to determine the structural and functional impact of novel missense variants on the keratin coiled-coil heterodimer. KVarPredDB was built under the integrative web application development framework SSM (SpringBoot, Spring MVC, MyBatis) and implemented in Java, Bootstrap, React-mutation-mapper, MySQL, Tomcat. The website can be accessed through http://bioinfo.zju.edu.cn/KVarPredDB . The genomic variants and analysis results are freely available under the Creative Commons license.

Conclusions: KVarPredDB provides an intuitive and user-friendly interface with computational analytical investigation for each missense variant of the keratin genes associated with genodermatoses.
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http://dx.doi.org/10.1186/s40246-020-00295-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7720490PMC
December 2020

The conserved microRNA miR-210 regulates lipid metabolism and photoreceptor maintenance in the Drosophila retina.

Cell Death Differ 2021 02 10;28(2):764-779. Epub 2020 Sep 10.

Division of Human Reproduction and Developmental Genetics, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310058, China.

Increasing evidence suggests that miRNAs play important regulatory roles in the nervous system. However, the molecular mechanisms of how specific miRNAs affect neuronal development and functions remain less well understood. In the present study, we provide evidence that the conserved microRNA miR-210 regulates lipid metabolism and prevents neurodegeneration in the Drosophila retina. miR-210 is specifically expressed in the photoreceptor neurons and other sensory organs. Genetic deletion of miR-210 leads to lipid droplet accumulation and photoreceptor degeneration in the retina. These effects are associated with abnormal activation of the Drosophila sterol regulatory element-binding protein signaling. We further identify the acetyl-coenzyme A synthetase (ACS) as one functionally important target of miR-210 in this context. Reduction of ACS in the miR-210 mutant background suppresses the neurodegeneration defects, suggesting that miR-210 acts through regulation of the ACS transcript. Together, these results reveal an unexpected role of miR-210 in controlling lipid metabolism and neuronal functions.
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http://dx.doi.org/10.1038/s41418-020-00622-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7862623PMC
February 2021

A motor neuron protective role of miR-969 mediated by the transcription factor kay.

RNA Biol 2020 09 13;17(9):1277-1283. Epub 2020 May 13.

Division of Human Reproduction and Developmental Genetics, Women's Hospital, Zhejiang University School of Medicine , Hangzhou, Zhejiang, China.

Maintenance of motor neuron structure and function is crucial in development and motor behaviour. However, the genetic regulatory mechanism of motor neuron function remains less well understood. In the present study, we identify a novel neuroprotective role of the microRNA miR-969 in motor neurons. miR-969 is highly expressed in motor neurons. Loss of miR-969 results in early-onset and age-progressive locomotion impairment. Flies lacking miR-969 also exhibit shortened lifespan. Moreover, miR-969 is required in motor neurons. We further identify kay as a functionally important target of miR-969. Together, our results indicate that miR-969 can protect motor neuron function by limiting kay activity in .
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http://dx.doi.org/10.1080/15476286.2020.1757897DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7549711PMC
September 2020

Prominin-like, a homolog of CD133, interacts with ND20 to maintain mitochondrial function.

Cell Biosci 2019 19;9:101. Epub 2019 Dec 19.

1College of Life Science, Zhejiang University, Hangzhou, 310058 Zhejiang China.

Background: Prominin-like is a homolog of mammalian CD133, which is recognized as a biomarker for stem cells. The interacting proteins of CD133 and their biological functions remain elusive.

Results: In this study, we using yeast two-hybrid assays, GST pull-down assay and co-immunoprecipitation (Co-IP) methods found that Prominin-like interacts with ND20, a subunit of mitochondrial respiratory complex I. Bioinformatics analysis suggests that Prominin-like is a six-transmembrane glycoprotein which localizes on cellular membranes. Immunostaining and mitochondrial fractionation indicate that Prominin-like could localize in the mitochondria. The knockdown of - in S2 cells resulted in transient mitochondrial dysfunctions as evidenced by reduced ATP production, elevated ROS generation and an accompanied reduction in mitochondrial proteins. Mitochondrial dysfunctions were detected in aged - mutant flies.

Conclusion: Our data indicates that Prominin-like acts to maintain mitochondrial function through its interaction with ND20 which, itself, is active in the mitochondrial electron transport chain. Our study provides insights into a novel molecular mechanism of - and suggests a similar function of CD133 in mammals.
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http://dx.doi.org/10.1186/s13578-019-0365-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6923988PMC
December 2019

A Novel Neuroprotective Role of Phosphatase of Regenerating Liver-1 against CO Stimulation in Drosophila.

iScience 2019 Sep 22;19:291-302. Epub 2019 Jul 22.

Institute of Genetics and Department of Genetics, Division of Human Reproduction and Developmental Genetics of the Women's Hospital, Zhejiang University School of Medicine, Yuhangtang Road 866, Xihu District, Hangzhou, Zhejiang Province 310058, China. Electronic address:

Neuroprotection is essential for the maintenance of normal physiological functions in the nervous system. This is especially true under stress conditions. Here, we demonstrate a novel protective function of PRL-1 against CO stimulation in Drosophila. In the absence of PRL-1, flies exhibit a permanent held-up wing phenotype upon CO exposure. Knockdown of the CO olfactory receptor, Gr21a, suppresses the phenotype. Our genetic data indicate that the wing phenotype is due to a neural dysfunction. PRL-1 physically interacts with Uex and controls Uex expression levels. Knockdown of Uex alone leads to a similar wing held-up phenotype to that of PRL-1 mutants. Uex acts downstream of PRL-1. Elevated Uex levels in PRL-1 mutants prevent the CO-induced phenotype. PRL-1 and Uex are required for a wide range of neurons to maintain neuroprotective functions. Expression of human homologs of PRL-1 could rescue the phenotype in Drosophila, suggesting a similar function in humans.
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http://dx.doi.org/10.1016/j.isci.2019.07.026DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6700421PMC
September 2019

The autophagy-related gene in regulates both neuron and midgut homeostasis.

J Biol Chem 2019 04 13;294(14):5666-5676. Epub 2019 Feb 13.

From the Division of Human Reproduction and Developmental Genetics, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China 310058,

is an autophagy-related gene identified in worms, flies, mice, and mammals, which encodes a protein that functions in autophagosome formation by associating with the ULK1-Atg13-Fip200 complex. In the last few years, the critical role of Atg101 in autophagy has been well-established through biochemical studies and the determination of its protein structure. However, Atg101's physiological role, both during development and in adulthood, remains less understood. Here, we describe the generation and characterization of an loss-of-function mutant in and report on the roles of Atg101 in maintaining tissue homeostasis in both adult brains and midguts. We observed that homozygous or hemizygous mutants were semi-lethal, with only some of them surviving into adulthood. Both developmental and starvation-induced autophagy processes were defective in the mutant animals, and mutant adult flies had a significantly shorter lifespan and displayed a mobility defect. Moreover, we observed the accumulation of ubiquitin-positive aggregates in mutant brains, indicating a neuronal defect. Interestingly, mutant adult midguts were shorter and thicker and exhibited abnormal morphology with enlarged enterocytes. Detailed analysis also revealed that the differentiation from intestinal stem cells to enterocytes was impaired in these midguts. Cell type-specific rescue experiments disclosed that Atg101 had a function in enterocytes and limited their growth. In summary, the results of our study indicate that Atg101 is essential for tissue homeostasis in both adult brains and midguts. We propose that Atg101 may have a role in age-related processes.
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http://dx.doi.org/10.1074/jbc.RA118.006069DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6462509PMC
April 2019

Drosophila Pif1A is essential for spermatogenesis and is the homolog of human CCDC157, a gene associated with idiopathic NOA.

Cell Death Dis 2019 02 11;10(2):125. Epub 2019 Feb 11.

Division of Human Reproduction and Developmental Genetics, the Women's Hospital, Zhejiang University School of Medicine, Yuhangtang Road 866, 310012, Hangzhou, Zhejiang, China.

The dynamic process of spermatogenesis shows little variation between invertebrate models such as Drosophila, and vertebrate models such as mice and rats. In each case, germ stem cells undergo mitotic division to proliferate and then continue, via meiosis, through various stages of elongation and individualization from spermatogonia to spermatid to finally to form mature sperm. Mature sperm are then stored in the seminal vesicles for fertilization. Errors in any of these stages can lead to male infertility. Here, we identify that Drosophila Pif1A acts as a key regulator for sperm individualization. Loss of Pif1A leads to male sterility associated with irregular individualization complex and empty seminal vesicles without mature sperm. Pif1A is highly expressed in the testes of mated male adult flies and the Pif1A protein is expressed at a higher level in male than in female flies. Pif1A is homologous to mammalian coiled-coil domain-containing protein 157 (CCDC157), which is also enriched in the testes of humans and mice. Human CCDC157, with unknown function, was identified to be downregulated in men with idiopathic non-obstructive azoospermia (NOA). We map the function of Drosophila Pif1A during spermatogenesis, showing that Pif1A is essential for spermatide individualization and involved in the regulation of the lipid metabolism genes. Our findings might be applicable for studying the function of CCDC157 in spermatogenesis and other aspects of human male fertility.
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http://dx.doi.org/10.1038/s41419-019-1398-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6370830PMC
February 2019

Hcf regulates the Hippo signaling pathway via association with the histone H3K4 methyltransferase Trr.

Biochem J 2019 02 28;476(4):759-768. Epub 2019 Feb 28.

Division of Human Reproduction and Developmental Genetics, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China

Control of organ size is a fundamental aspect in biology and plays important roles in development. The Hippo pathway is a conserved signaling cascade that controls tissue and organ size through the regulation of cell proliferation and apoptosis. Here, we report on the roles of Hcf (host cell factor), the homolog of Host cell factor 1, in regulating the Hippo signaling pathway. Loss-of-Hcf function causes tissue undergrowth and the down-regulation of Hippo target gene expression. Genetic analysis reveals that Hcf is required for Hippo pathway-mediated overgrowth. Mechanistically, we show that Hcf associates with the histone H3 lysine-4 methyltransferase Trithorax-related (Trr) to maintain H3K4 mono- and trimethylation. Thus, we conclude that Hcf positively regulates Hippo pathway activity through forming a complex with Trr and controlling H3K4 methylation.
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http://dx.doi.org/10.1042/BCJ20180717DOI Listing
February 2019

RanGAP-mediated nucleocytoplasmic transport of Prospero regulates neural stem cell lifespan in Drosophila larval central brain.

Aging Cell 2019 02 13;18(1):e12854. Epub 2018 Dec 13.

Division of Human Reproduction and Developmental Genetics, The Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China.

By the end of neurogenesis in Drosophila pupal brain neuroblasts (NBs), nuclear Prospero (Pros) triggers cell cycle exit and terminates NB lifespan. Here, we reveal that in larval brain NBs, an intrinsic mechanism facilitates import and export of Pros across the nuclear envelope via a Ran-mediated nucleocytoplasmic transport system. In rangap mutants, the export of Pros from the nucleus to cytoplasm is impaired and the nucleocytoplasmic transport of Pros becomes one-way traffic, causing an early accumulation of Pros in the nuclei of the larval central brain NBs. This nuclear Pros retention initiates NB cell cycle exit and leads to a premature decrease of total NB numbers. Our data indicate that RanGAP plays a crucial role in this intrinsic mechanism that controls NB lifespan during neurogenesis. Our study may provide insights into understanding the lifespan of neural stem cells during neurogenesis in other organisms.
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http://dx.doi.org/10.1111/acel.12854DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6351831PMC
February 2019

A positive role of Sin3A in regulating Notch signaling during Drosophila wing development.

Cell Signal 2019 01 12;53:184-189. Epub 2018 Oct 12.

Division of Human Reproduction and Developmental Genetics, The Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China; Institute of Genetics, Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China. Electronic address:

Notch is a transmembrane receptor that mediates intercellular signaling through a conserved signaling cascade in all animal species. Transcriptional and posttranscriptional regulation of Notch receptor are important for maintaining Notch signaling activity. Here, we show that depletion of Drosophila Sin3A leads to loss of the adult wing margin and downregulation of Notch target gene expression in the developing wing disc. Sin3A regulates the Notch pathway downstream of Delta and upstream of Notch activation. The role of Sin3A in the Notch pathway is partly mediated by its ability to modulate Notch receptor transcription. Furthermore, the transcriptional activation of Notch receptor is autoregulated by Notch itself. We also provide evidence that Sin3A is required for Notch activation mediated Notch transcription. Together, our data demonstrate that Sin3A activates Notch signaling by promoting Notch transcription and reveal a previously unknown autoregulatory mechanism for Notch signaling activation during Drosophila wing development.
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http://dx.doi.org/10.1016/j.cellsig.2018.10.008DOI Listing
January 2019

Prominin-like, a homolog of mammalian CD133, suppresses di lp6 and TOR signaling to maintain body size and weight in Drosophila.

FASEB J 2019 02 11;33(2):2646-2658. Epub 2018 Oct 11.

Division of Human Reproduction and Developmental Genetics, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, China.

CD133 (AC133/prominin-1) has been identified as a stem cell marker and a putative cancer stem cell marker in many solid tumors. Its biologic function and molecular mechanisms remain largely elusive. Here, we show that a fly mutant for prominin-like, a homolog of mammalian CD133, shows a larger body size and excess weight accompanied with higher fat deposits as compared with the wild type. The expression levels of prominin-like are mediated by ecdysone signaling where its protein levels increase dramatically in the fat body during metamorphosis. Prominin-like mutants exhibit higher Drosophila insulin-like peptide 6 (di lp6) levels during nonfeeding stages and increased Akt/ Drosophila target of rapamycin (dTOR) signaling. On an amino acid-restricted diet, prominin-like mutants exhibit a significantly larger body size than the wild type does, similar to that which occurs upon the activation of the dTOR pathway in the fat body. Our data suggest that prominin-like functions by suppressing TOR and dilp6 signaling to control body size and weight. The identification of the physiologic function of prominin-like in Drosophila may provide valuable insight into the understanding of the metabolic function of CD133 in mammals.-Zheng, H., Zhang, Y., Chen, Y., Guo, P., Wang, X., Yuan, X., Ge, W., Yang, R., Yan, Q., Yang, X., Xi, Y. Prominin-like, a homolog of mammalian CD133, suppresses di lp6 and TOR signaling to maintain body size and weight in Drosophila.
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http://dx.doi.org/10.1096/fj.201800123RDOI Listing
February 2019

The histone deacetylase HDAC1 positively regulates Notch signaling during wing development.

Biol Open 2018 Feb 20;7(2). Epub 2018 Feb 20.

Division of Human Reproduction and Developmental Genetics, The Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China

The Notch signaling pathway is highly conserved across different animal species and plays crucial roles in development and physiology. Regulation of Notch signaling occurs at multiple levels in different tissues and cell types. Here, we show that the histone deacetylase HDAC1 acts as a positive regulator of Notch signaling during wing development. Depletion of causes wing notches on the margin of adult wing. Consistently, the expression of Notch target genes is reduced in the absence of HDAC1 during wing margin formation. We further provide evidence that HDAC1 acts upstream of Notch activation. Mechanistically, we show that HDAC1 regulates Notch protein levels by promoting Notch transcription. Consistent with this, the HDAC1-associated transcriptional co-repressor Atrophin (Atro) is also required for transcriptional activation of Notch in the wing disc. In summary, our results demonstrate that HDAC1 positively regulates Notch signaling and reveal a previously unidentified function of HDAC1 in Notch signaling.
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http://dx.doi.org/10.1242/bio.029637DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5861358PMC
February 2018

Real-time observation of perturbation of a Drosophila embryo's early cleavage cycles with microfluidics.

Anal Chim Acta 2017 Aug 13;982:131-137. Epub 2017 Jun 13.

Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou 310058, China. Electronic address:

It is of great importance to understand biochemical system's behavior toward environmental perturbation during the development of living organisms. Here a microfluidic platform for Drosophila embryo's online development and observation is presented. The system is capable of developing the embryo's anterior and posterior halves controlled at different temperature environments, and it can be easily coupled with a confocal microscope for real-time image acquisition. The microfluidic chip is consisted of a polymethylmethacrylate (PMMA) substrate with a thickness of 4.0 mm and a polydimethylsiloxane (PDMS) cover designed with a typical 'Y' channel with a depth of 400 μm, width of 800 μm. Temperature gradients were created across the anterior half and posterior half of the embryo by utilizing two streams of laminar flow with different temperatures. It was found that thermal gradient would result in asynchronous development of the two halves of the embryos, and the developing difference was related to the direction of thermal gradient. This may result from the presence of an unknown mechanism located in the anterior half of the embryo, which oversees nuclear division synchronicity. These observations would help better understand compensatory mechanisms of Drosophila embryo's development under environmental perturbations.
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http://dx.doi.org/10.1016/j.aca.2017.05.024DOI Listing
August 2017

Inscuteable maintains type I neuroblast lineage identity via Numb/Notch signaling in the Drosophila larval brain.

J Genet Genomics 2017 03 6;44(3):151-162. Epub 2017 Mar 6.

Division of Human Reproduction and Developmental Genetics, The Women's Hospital, School of Medicine, Zhejiang University, Hangzhou 310000, China; Institute of Genetics, Zhejiang University, Hangzhou 310012, China; Department of Genetics, School of Medicine, Zhejiang University, Hangzhou 310012, China; Joint Institute of Genetics and Genomic Medicine Between Zhejiang University and University of Toronto, Zhejiang University, Hangzhou 310012, China. Electronic address:

In the Drosophila larval brain, type I and type II neuroblasts (NBs) undergo a series of asymmetric divisions which give rise to distinct progeny lineages. The intermediate neural progenitors (INPs) exist only in type II NB lineages. In this study, we reveal a novel function of Inscuteable (Insc) that acts to maintain type I NB lineage identity. In insc type I NB clones of mosaic analyses with a repressible cell marker (MARCM), the formation of extra Deadpan (Dpn) NB-like and GMC-like cells is observed. The lack of Insc leads to the defective localization and segregation of Numb during asymmetric cell division. By the end of cytokinesis, this results in insufficient Numb in ganglion mother cells (GMCs). The formation of extra Deadpan (Dpn) cells in insc clones is prevented by the attenuation of Notch activity. This suggests that Insc functions through the Numb/Notch signaling pathway. We also show that in the absence of Insc in type I NB lineages, the cellular identity of GMCs is altered where they adopt an INP-like cell fate as indicated by the initiation of Dpn expression accompanied by a transient presence of Earmuff (Erm). These INP-like cells have the capacity to divide multiple times. We conclude that Insc is necessary for the maintenance of type I NB lineage identity. Genetic manipulations to eliminate most type I NBs with overproliferating type II NBs in the larval brain lead to altered circadian rhythms and defective phototaxis in adult flies. This indicates that the homeogenesis of NB lineages is important for the adult's brain function.
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http://dx.doi.org/10.1016/j.jgg.2017.02.005DOI Listing
March 2017

Premature remodeling of fat body and fat mobilization triggered by platelet-derived growth factor/VEGF receptor in .

FASEB J 2017 05 26;31(5):1964-1975. Epub 2017 Jan 26.

Division of Human Reproduction and Developmental Genetics, The Women's Hospital, and

In fat-body remodeling accompanied with fat mobilization is an ecdysone-induced dynamic process that only occurs during metamorphosis. Here, we show that the activated platelet-derived growth factor/VEGF receptor (PVR) is sufficient to induce shape changes in the fat body, from thin layers of tightly conjugated polygonal cells to clusters of disaggregated round-shaped cells. These morphologic changes are reminiscent of those seen during early pupation upon initiation of fat-body remodeling. Activation of PVR also triggers an early onset of lipolysis and mobilization of internal storage, as revealed by the appearance of small lipid droplets and up-regulated lipolysis-related genes. We found that PVR displays a dynamic expression pattern in the fat body and peaks at the larval-prepupal transition under the control of ecdysone signaling. Removal of PVR, although it does not prevent ecdysone-induced fat-body remodeling, causes ecdysone signaling to be up-regulated. Our data reveal that PVR is active in a dual-secured mechanism that involves an ecdysone-induced fat-body remodeling pathway and a reinforced PVR pathway for effective lipid mobilization. Ectopic expression of activated c-kit-the mouse homolog of PVR in the fat body-also results in a similar phenotype. This may suggest a novel function of c-kit as it relates to lipid metabolism in mammals.-Zheng, H., Wang, X., Guo, P., Ge, W., Yan, Q., Gao, W., Xi, Y., Yang, X. Premature remodeling of fat body and fat mobilization triggered by platelet-derived growth factor/VEGF receptor in .
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http://dx.doi.org/10.1096/fj.201601127RDOI Listing
May 2017

Fat body remodeling and homeostasis control in Drosophila.

Life Sci 2016 Dec 20;167:22-31. Epub 2016 Oct 20.

Division of Human Reproduction and Developmental Genetics, the Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China. Electronic address:

Remarkable advances have been made in recent years in our understanding of the Drosophila fat body and its functions in energy storage, immune response and nutrient sensing. The fat body interplays with other tissues to respond to the physiological needs of the body's growth and coordinates various metabolic processes at different developmental stages and under different environmental conditions. The identification of various conserved genetic functions and signaling pathways relating to the Drosophila fat body may provide clues to lipometabolic disease and other aspects of tissue remodeling in humans. Here, we discuss recent insights into how regulation of fat body remodeling contributes to hemostasis with a special focus on how signaling networks and internal physiological states shape different aspects of the lipid metabolism in Drosophila.
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http://dx.doi.org/10.1016/j.lfs.2016.10.019DOI Listing
December 2016

Cross-Talk Between Mitochondrial Fusion and the Hippo Pathway in Controlling Cell Proliferation During Drosophila Development.

Genetics 2016 08 17;203(4):1777-88. Epub 2016 Jun 17.

Division of Human Reproduction and Developmental Genetics, The Women's Hospital, Zhejiang University School of Medicine, Hangzhou, China 310058 Institute of Genetics and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, China 310058

Cell proliferation and tissue growth depend on the coordinated regulation of multiple signaling molecules and pathways during animal development. Previous studies have linked mitochondrial function and the Hippo signaling pathway in growth control. However, the underlying molecular mechanisms are not fully understood. Here we identify a Drosophila mitochondrial inner membrane protein ChChd3 as a novel regulator for tissue growth. Loss of ChChd3 leads to tissue undergrowth and cell proliferation defects. ChChd3 is required for mitochondrial fusion and removal of ChChd3 increases mitochondrial fragmentation. ChChd3 is another mitochondrial target of the Hippo pathway, although it is only partially required for Hippo pathway-mediated overgrowth. Interestingly, lack of ChChd3 leads to inactivation of Hippo activity under normal development, which is also dependent on the transcriptional coactivator Yorkie (Yki). Furthermore, loss of ChChd3 induces oxidative stress and activates the JNK pathway. In addition, depletion of other mitochondrial fusion components, Opa1 or Marf, inactivates the Hippo pathway as well. Taken together, we propose that there is a cross-talk between mitochondrial fusion and the Hippo pathway, which is essential in controlling cell proliferation and tissue homeostasis in Drosophila.
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http://dx.doi.org/10.1534/genetics.115.186445DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4981277PMC
August 2016

Phosphotyrosyl phosphatase activator facilitates localization of Miranda through dephosphorylation in dividing neuroblasts.

Development 2016 Jan 19;143(1):35-44. Epub 2015 Nov 19.

College of Life Sciences, Zhejiang University, Hangzhou 310058, China Institute of Genetics, School of Medicine, Zhejiang University, Hangzhou 310058, China

The mechanism for the basal targeting of the Miranda (Mira) complex during the asymmetric division of Drosophila neuroblasts (NBs) is yet to be fully understood. We have identified conserved Phosphotyrosyl phosphatase activator (PTPA) as a novel mediator for the basal localization of the Mira complex in larval brain NBs. In mutant Ptpa NBs, Mira remains cytoplasmic during early mitosis and its basal localization is delayed until anaphase. Detailed analyses indicate that PTPA acts independent of and before aPKC to localize Mira. Mechanistically, our data show that the phosphorylation status of the T591 residue determines the subcellular localization of Mira and that PTPA facilitates the dephosphorylation of T591. Furthermore, PTPA associates with the Protein phosphatase 4 complex to mediate localization of Mira. On the basis of these results, a two-step process for the basal localization of Mira during NB division is revealed: cortical association of Mira mediated by the PTPA-PP4 complex is followed by apical aPKC-mediated basal restriction.
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http://dx.doi.org/10.1242/dev.127233DOI Listing
January 2016

Alterations of circadian clockworks during differentiation and apoptosis of rat ovarian cells.

Chronobiol Int 2011 Jul;28(6):477-87

Department of Animal and Marine Bioresource Sciences, Graduate School of Agriculture, Kyushu University, Fukuoka, Japan.

Ovarian development is related to cell proliferation, differentiation, and apoptosis of granulosa cells and luteal cells under the control of various modulators, including follicle-stimulating hormone (FSH), luteinizing hormone (LH), and growth factors. In the present study, the expression of clock genes and the related regulation mechanism were analyzed in different ovarian cell types during differentiation and apoptosis. The authors focused on the circadian expression of Per2 as a core clock gene for the maintenance of circadian rhythms. By using a real-time monitoring system of the Per2 promoter activity, the circadian oscillation was analyzed in the granulosa and luteal cells from preantral follicles, antral follicles, and corpora lutea of immature Per2 promoter-destabilized luciferase transgenic rats that were primed with diethylstilbestrol, equine chorionic gonadotropin (eCG), and/or human CG. In addition, transcript levels of Per2, Bmal1, Clock, and Nampt were quantified by quantitative polymerase chain reaction (qPCR). Immunohistochemical studies revealed strong circadian rhythmicity of PER2 protein in the luteal cells, but apparently little rhythmicity in granulosa cells of both preantral and antral follicles. In vitro monitoring of promoter activity showed generation of several oscillations in luteal cells after exposure to dexamethasone (DXM), whereas oscillatory amplitudes of immature and mature granulosa cells were rapidly attenuating. The circadian rhythm of the Bmal1 transcript levels, but not the Per2 transcript, was very weak in the granulosa cells, as compared with that in luteal cells. Granulosa cells gained a strong circadian rhythm ability of the Per2 promoter activity after stimulation with FSH for 3 days. In contrast, LH had little effect on the circadian rhythm before stimulation of granulosa cells with FSH, probably owing to lack of LH receptor. In luteal cells, induction of apoptosis by inhibiting progesterone synthesis resulted in deregulation of Per2 circadian oscillation. Transcript levels of Bmal1 and Clock, but not Per2 and Nampt, were significantly decreased in apoptotic luteal cells. The Bmal1 transcript level was particularly reduced. Consequently, these results strongly suggest the circadian clockwork alters in ovarian cells during follicular development, luteinization, and apoptosis, and expression of Bmal1 may be related to the switch-on and switch-off of the circadian oscillation.
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http://dx.doi.org/10.3109/07420528.2011.589933DOI Listing
July 2011

Effects of ectopic expression of human telomerase reverse transcriptase on immortalization of feather keratinocyte stem cells.

J Exp Zool B Mol Dev Evol 2009 Dec;312(8):872-84

Institute of Cell Biology and Genetics, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang Province, China.

Normal somatic cells possess a finite life span owing to replicative senescence. Telomerase functions as a potential regulator of senescence in various cells. Expression level of human telomerase reverse transcriptase (hTERT) is correlated with telomerase activity and cellular immortalization. In this study, we investigated the effects of ectopic expression of hTERT on proliferation potential of chicken feather keratinocyte stem cells (FKSCs). We established FKSCs transduced with hTERT catalytic subunit fused with EGFP marker gene (hTERT-EGFP-FKSCs). hTERT-EGFP-FKSCs had the great potential of proliferation in vitro and expressed kerainocyte stem cell markers integrin beta1 and CD49c. Keratin 15 and keratin 19, as native FKSCs, were also detected in hTERT-EGFP-FKSCs. By the analysis of fluorescent RT-PCR, western blotting and TRAP assay, hTERT-EGFP-FKSCs were positive for telomerase activity, in comparison with native FKSCs showing no telomerase activity. We demonstrated that ectopic expression of hTERT could result in immortalization of FKSCs. Tumorigenecity of hTERT-EGFP-FKSCs were examined by soft agar assay and transplantation into NOD-SCID mice. Results showed that hTERT-EGFP-FKSCs sustained the cellular characteristics of native FKSCs and had no transforming activity. In vivo differentiation multipotentials of hTERT-EGFP-FKSCs were confirmed by transplantation into developing chicken embryos and in situ hybridization analysis. These data provide a novel framework for understanding human telomerase activity in different species and suggest a new insight for manipulating hTERT for therapeutic purposes in treating tissue injury and aging.
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http://dx.doi.org/10.1002/jez.b.21302DOI Listing
December 2009

Effects of hindlimb unloading on ex vivo growth and osteogenic/adipogenic potentials of bone marrow-derived mesenchymal stem cells in rats.

Stem Cells Dev 2008 Aug;17(4):795-804

College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.

The goal of this study was to determine the effects of hindlimb unloading (HU) on the ex vivo growth and the osteogenic potential of mesenchymal stem cells (MSCs) from the femurs of rats. Microgravity was simulated by 28-day HU in male Sprague-Dawley (SD) rats, and the bone marrow (BM) was collected from hindlimb femurs of HU or control (CTL) rats. MSCs were isolated from BM and cultured for eight passages. Then MSCs at passages 2, 4, and 8 were induced for osteogenesis or adipogenesis. The results revealed that HU decreased the osteogenic potential of MSCs and also decreased the expression of osteoblast gene marker mRNAs in cells induced by osteogenic conditions. Meanwhile, the expression of Runx2 mRNA and the phosphorylation of ERK were also decreased. There were no significant differences of osteoblast gene marker and Runx2 mRNA expression between cells induced from different passages of MSCs in UH rats. Under adipogenic conditions, HU increased both the adipogenic potential of MSCs and the expression of adipocytic gene marker mRNAs in induced cells. HU also increased the expression of PPAR gamma 2 mRNA, but with no effect on the phosphorylation of p38MAPK. The adipogenic potential of MSCs and the expression of adipocytic gene marker mRNAs in induced cells decreased along with cell cultures under normal gravity. This suggests that the normal gravity during in vitro MSC culture and the centrifugal force produced during cell harvest after each passage could decrease the adipogenic potential of MSCs, but could not reverse the effect of HU on the osteogenic potential of MSCs.
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http://dx.doi.org/10.1089/scd.2007.0254DOI Listing
August 2008

Prevalence of a septicemia disease in the crested ibis (Nipponia nippon) in China.

Avian Dis 2007 Jun;51(2):614-7

The Institute of Genetics and Cell Biology, College of Life Sciences, Zhejiang University, 568 Zijinhua Road, Hangzhou, Zhejiang Province, 310058, People's Republic of China.

This study investigated six cases of septicemia in young crested ibises (Nipponia nippon). These birds all died with similar clinical signs, including sudden death, anorexia, diarrhea, and lameness. Immediately after death, the birds were necropsied; a blood sample was taken from heart and tissues were sampled from liver, lung, spleen, peritoneal mucus, and feces for bacteriologic examination. Anatomic observation showed that the main findings common to the sick birds were arthrocele, associated with congestion in the femur, tibiotarsus, and ventral side; swelling in the liver; hemorrhagic pericarditis; miliary tubercles in lung; and fibrous tubercles in the synovial capsule of the knee joint with suppurative abscesses. Through bacterial examination, the colonial type of Escherichia coli strain was represented prominently in cultures of the feces, heart blood, liver, lung, spleen, suppurative mucus of the synovial capsule, and peritoneal exudate. These symptoms suggested that the death of a number of endangered crested ibis within a short period was evidence of septicemia. The bacterial inoculation tests were also conducted using domestic pigeon, native chicken, and mice for the presence of and infection with E coli. The study provided indications of the possible role of E. coli strains as bird pathogens and a potential risk in endangered species. Further work is needed to characterize E. coli strains and the toxin production in this bird. This disease occurrence also adds a note of caution to the continued efforts and interest in the reintroduction of the ibis back into its former wild ranges to ensure that formerly captive individuals do not transmit disease to the wild populations of its own or other sympatric species.
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http://dx.doi.org/10.1637/0005-2086(2007)51[614:POASDI]2.0.CO;2DOI Listing
June 2007

Could the effect of modeled microgravity on osteogenic differentiation of human mesenchymal stem cells be reversed by regulation of signaling pathways?

Biol Chem 2007 Jul;388(7):755-63

Department of Cell Biology, College of Life Science, Zhejiang University, Hangzhou, Zhejiang, PR China.

Microgravity (MG) results in a reduction in bone formation. Bone formation involves osteogenic differentiation from mesenchymal stem cells (hMSCs) in bone marrow. We modeled MG to determine its effects on osteogenesis of hMSCs and used activators or inhibitors of signaling factors to regulate osteogenic differentiation. Under osteogenic induction, MG reduced osteogenic differentiation of hMSCs and decreased the expression of osteoblast gene markers. The expression of Runx2 was also inhibited, whereas the expression of PPARgamma2 increased. MG also decreased phosphorylation of ERK, but increased phosphorylation of p38MAPK. SB203580, a p38MAPK inhibitor, was able to inhibit the phosphorylation of p38MAPK, but did not reduce the expression of PPARgamma2. Bone morphogenetic protein (BMP) increased the expression of Runx2. Fibroblast growth factor 2 (FGF2) increased the phosphorylation of ERK, but did not significantly increase the expression of osteoblast gene markers. The combination of BMP, FGF2 and SB203580 significantly reversed the effect of MG on osteogenic differentiation of hMSCs. Our results suggest that modeled MG inhibits the osteogenic differentiation and increases the adipogenic differentiation of hMSCs through different signaling pathways. Therefore, the effect of MG on the differentiation of hMSCs could be reversed by the mediation of signaling pathways.
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http://dx.doi.org/10.1515/BC.2007.082DOI Listing
July 2007

Establishment of feather follicle stem cells as potential vehicles for delivering exogenous genes in birds.

J Reprod Dev 2003 Jun;49(3):213-9

Laboratory of Reproductive Physiology and Biotechnology, Department of Animal and Marine Bioresource Sciences, Faculty of Agriculture, Graduate School Kyushu University, Fukuoka, Japan.

The present study was performed to develop a culture system for feather keratinocyte stem cells to enable the genetic manipulation of endangered avian species. The feather follicle cells were isolated from growing feathers of adult White Leghorn chicken. Leukemia inhibitory factor (LIF) was used to maintain the characterization of the keratinocyte colony-forming cells (KCFCs). The EGFPN1 plasmid DNA retroviral vector was used to deliver Green Fluorescent Protein (GFP) gene, which was introduced to the KCFCs by lipofection. After removal of the fibroblast-like cells, the feather KCFCs attached to the substrate within 24 h of seeding. The cells continued to proliferate for at least 30 days in the presence of LIF. The cell-adhesion molecules such as integrin beta1 and CD49c were immunocytochemically positive in the cells. The KCFCs differentiated into barbular cells and pennaceous feather vane in the LIF-free medium. The GFP gene-transfected KCFCs stably expressed GFP. The present results indicate that the KCFCs derived from feather follicles are closely related to multipotent stem cells. In addition, gene manipulation of such stem cells may be useful for the production of chimera in avian species.
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http://dx.doi.org/10.1262/jrd.49.213DOI Listing
June 2003

Green fluorescent protein gene-transfected peafowl somatic cells participate in the development of chicken embryos.

J Exp Zool A Comp Exp Biol 2004 Feb;301(2):139-49

Laboratory of Reproductive Physiology and Biotechnology, Department of Animal and Marine Bioresource Sciences, Graduate School Kyushu University, Fukuoka 812-8581, Japan.

This study was performed to investigate whether the embryonic somatic cells are capable of reconstituting and participating in the embryonic development of chickens to produce chimeras. In order to track the migration behavior of the donor cells, a cell line, originally isolated from an Indian peafowl embryo, was fluorescent-labeled by transfection of the cells with enhanced Green Fluorescent Protein (GFP) and Neomycin resistant (Neo) genes prior to injection into the stage X blastoderm of White Leghorn chickens. The injection was performed with a medium in the presence of 1-5% polyethylene glycol. The development of putative chimeric embryos between the stages three and 24 was examined for GFP expression under fluorescent light. To trace the peafowl cells in the developing chicken embryos, both a species-specific genetic marker originating from the mitochondrial DNA cytochrome b (cyt b) gene and a DNA fragment of GFP gene were used. Of the 185 fertile eggs manipulated, 173 developed into embryos. Fifty-five of them showed positive GFP patches in extra-embryonic tissues, and 15 expressed GFP in intra-embryonic tissues such as those of the head, heart, and gonad. PCR analysis revealed that PCR fragments for the peafowl mitochondrial DNA cyt b and GFP genes were detected in the samples of the GFP positive extra- and intra-embryonic tissues of the chimeras. The present results provide evidence that fluorescent-labeled peafowl embryonic cells carrying GFP and Neo genes are able to participate in the development of chicken embryos to generate chimeras.
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http://dx.doi.org/10.1002/jez.a.20021DOI Listing
February 2004
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